ABSTRACT
The present study investigated the effect of ivermectin and amitraz on the cellular architecture of vital organs of Rhipicephalus microplus. Adult female ticks were treated with lethal concentrations (LC95) of ivermectin and amitraz, and the ovaries, synganglion, and Gené's organ were processed 48 h post treatment. In both the treatment groups, the ultra-thin sections of ovary exhibited deformed oocytes, irregular plasmic membrane and chorion layer, extensive vacuolation in the cytoplasm mainly at periphery of the cell and oocyte-pedicel junction. Marked vacuolations in the cortex and neuropile region with significant structural disorganization of the neural fibers were common alterations observed in the synganglion of ticks exposed to ivermectin and amitraz. The tissue sections of Gené's organ revealed deformed tubular glands with severe loss of cellular limit of secretory epithelium and cytoplasmic vacuolations in the ivermectin treated ticks whereas, the alterations were comparatively less severe in amitraz exposed ticks. The cellular deformities in these vital organs probably impaired reproductive function, nerve signal transmission and metabolic activities and thus affected fecundity and survivability of the treated ticks. The findings suggested that the action of ivermectin and amitraz are not restricted to the nervous system of ticks, but also on other vital organs, ovary and Gené's organ affecting the oviposition. The study provided insights into the development of targeted interventions for tick control strategies.
Subject(s)
Ivermectin , Ovary , Rhipicephalus , Toluidines , Animals , Ivermectin/pharmacology , Female , Rhipicephalus/drug effects , Toluidines/pharmacology , Ovary/drug effects , Acaricides/pharmacology , Microscopy, Electron, TransmissionABSTRACT
Canine monocytic ehrlichiosis caused by Ehrlichia canis is an important rickettsial pathogen of dogs transmitted by Rhipicephalus sanguineus sensu lato ticks in India. Globally, molecular characterization of E. canis is done using different E. canis gene targets. This study aimed to characterize genetic diversity of uncultured Ehrlichia species from dogs by 16S rRNA and partial gp200 gene (termed as p43 region) sequences data. Phylogeny based on 16S rRNA gene did not reveal any region-specific lineages. The phylogeny based on 5' region of E. canis gp200 gene (termed as p43 region) revealed four major clusters (A, B, C and D) and the Indian isolates fall under clusters A and B. Cluster A is characterized by an insertion of unique 141 bp tandem repeat sequence. Similar tandem repeat sequence was found in one of the E canis isolates from east-Asia, suggesting a possible divergence within this species. The study shows evidence for divergence of a new lineage within E. canis. The location of this insertion at the 'ankyrin repeat domains' containing region is suggestive of its possible role in modulation of host responses.
Subject(s)
Dog Diseases , Ehrlichia canis , Ehrlichiosis , Phylogeny , RNA, Ribosomal, 16S , Animals , Dogs , Ehrlichia canis/genetics , Ehrlichia canis/isolation & purification , Dog Diseases/microbiology , India , Ehrlichiosis/veterinary , Ehrlichiosis/microbiology , Ehrlichiosis/epidemiology , RNA, Ribosomal, 16S/genetics , Genetic Variation , DNA, Bacterial/geneticsABSTRACT
Ticks of the genus Rhipicephalus infesting cattle are the primary animal pests responsible for the annual economic loss of billions of dollars. Due to the morphological resemblance among the members of the Rhipicephalus (Boophilus) genus, species identification is very difficult. In this study, the adult R. annulatus and R. microplus ticks from two south Indian states viz., Kerala and Karnataka were subjected to morphological and molecular characterization. The R. microplus isolates from south India differed morphologically from true R. microplus clade A ticks. The ventral spur on the first pedipalp observed in male R. microplus was similar to that of R. australis. The phylogenetic analysis revealed that the R. microplus from these states clustered with R. microplus clade C. The mitochondrial cytochrome c oxidase I (COI) was identified as the preferred molecular marker compared to the internal transcribed spacer 2 (ITS2). The interspecific divergence between R. microplus and R. annulatus isolates from South India was 7.9 per cent based on COI. Moreover, based on COI, the R. microplus isolates revealed higher intraspecific divergence (2.9%) than R. annulatus (1%). The ITS2 sequences failed to differentiate R. microplus and R. annulatus.
Subject(s)
Cattle Diseases , Rhipicephalus , Tick Infestations , Male , Animals , Cattle , Rhipicephalus/anatomy & histology , Phylogeny , India , Cattle Diseases/epidemiology , Tick Infestations/epidemiology , Tick Infestations/veterinaryABSTRACT
As per 20th livestock census, India possessing 193.46 million cattle and 109.85 million buffaloes in organized and unorganized sectors. These animals are suffering from tick infestation almost throughout the year causing both direct and indirect losses. Repeated use of chemical acaricides for tick management resulted in the establishment of acaricide resistant tick populations, insecticide residues in livestock products and environmental pollution. In the present study, analysis of the data generated during 1990-2021 revealed pooled prevalence of infestations in cattle and buffaloes was 53% [95% CI = 47-60%] and 38% [95% CI = 28-49%], respectively. Based on peripheral blood smear examination the prevalence of theileriosis, babesiosis and anaplasmosis in cattle is estimated as 17% [95% CI = 12-24%], 4% [95% CI = 3-6%] and 5% [95% CI = 3-7%], respectively, while in buffalo the prevalence of infection is estimated as 7% [95% CI = 2-21%], 2% [95% CI = 0-5%] and 8% [95% CI = 2-36%],respectively.To estimate economic impact, both direct (reduction in milk production, cost of treatment, leather depreciation) and indirect (milk loss and treatment cost) losses were taken into consideration. Loss of milk production was predicted as 13.91, 56.91 and 85.34L/cross-bred cow/lactation in low, moderate and high tick infestation conditions, respectively. Whereas, 20.10, 7.01L milk/buffalo/lactation in Hyalomma spp. and Rhipicephalus spp. infestation was estimated. Similarly,the estimated loss of milk production due to clinical theileriosis, babesiosis and anaplasmosis was 57.96, 30.96 and 59.22L, respectively. The cumulative (milk loss, treatment cost and leather loss) loss due to tick infestation was calculated as 46199.31 million INR (USD595.07 million) while due to TBDs 14877.15 million INR (USD191.15 million) = 61076.46 million INR (USD787.63 million). The data provided base line information for the policy maker to develop strategies at government level so that the significantly high cumulative loss of 787.63 million USD due to ticks and tick borne diseases (TTBDs) can be minimized.
Subject(s)
Acaricides , Anaplasmosis , Babesiosis , Cattle Diseases , Rhipicephalus , Theileriasis , Tick Infestations , Tick-Borne Diseases , Animals , Female , Cattle , Tick Infestations/epidemiology , Tick Infestations/veterinary , Theileriasis/epidemiology , Buffaloes , Cattle Diseases/epidemiology , Tick-Borne Diseases/epidemiology , Tick-Borne Diseases/veterinary , LivestockABSTRACT
The present study was taken up to evaluate the synergistic properties of piperonyl butoxide (PBO), diethyl maleate (DEM), triphenyl phosphate (TPP) and verapamil (VER) with deltamethrin (DLM) and ivermectin (IVM) against DLM and IVM resistant tick populations collected from Madhya Pradesh and Punjab states of India. The collected field tick populations were resistant to DLM (Resistance Factor [RF] in the range of 21.71-32.98) and IVM (RF in the range of 1.89-4.98). A strong synergism between DLM and, IVM with PBO and IVM with VER was noticed. The synergistic efficacy of PBO and VER with IVM in reducing the lethal concentration 50 (LC50) value (1.69-5.72 times for PBO and 3.00-10.62 times for VER) of IVM in resistant ticks suggest that a combination of these synergists with IVM can significantly enhance the effectiveness of IVM against IVM-resistant Rhipicephlaus microplus populations gradually establishing in the different parts of the country. The synergistic efficiency of PBO with DLM in reducing the LC50 value was 2.65 and 18.01 times, respectively, against DLM- resistant two R. microplus populations (KTN and LDH). The study revealed the gradual establishment of DLM and IVM resistant populations in the surveyed states suggesting the need to adopt required resistance management strategies. The use of synergists with DLM and IVM has emerged as an effective approach for controlling the acaricide-resistant ticks.
ABSTRACT
Chemical acaricides are widely used for the effective control of ticks in India. The synthetic pyrethroids, are one of the most popular chemical acaricides with selective neurotoxic potential. Flumethrin is a type II synthetic pyrethroid used extensively in veterinary practice in India. The present study was undertaken to evaluate the cytotoxic effects of flumethrin on the engorged females of Rhipicephalus annulatus using entomological parameters, histology, electron microscopy and relative quantification of receptors of dopamine and GABAB mRNAs. Adult immersion test (AIT) using flumethrin (100 ppm), revealed twenty per cent mortality of ticks, hundred per cent inhibition of fecundity and complete blocking of hatching of the laid eggs. Microscopic analysis of the structure of the ovaries after 24 h of treatment with flumethrin (90 ppm) revealed changes, viz., reduction in size with the presence of amorphous material inside stage I oocytes, wrinkled boundary and chromatin fragmentation of nucleus of stage II oocytes, vacuoles around the germinal vesicle, thickening of the nuclear membrane and chromatin clumping of stage III oocytes and reduction in size and shape of mature stage IV and V oocytes. Also, a large number of vacuoles were observed throughout the pedicel cell region of stage II and III oocytes. Ultrastructurally, irregular nuclear membrane, swelling as well as crystolysis of mitochondria and detachment of external and internal layers of the basal lamina of oocytes were the major structural alterations confirming direct damaging effects of flumethrin on the germinative cells. The relative quantification of the expression of dopamine D1, dopamine D2 and GABAB receptors by quantitative real-time PCR (qRT PCR), revealed the upregulation of dopamine D1 receptor and downregulation of receptors of dopamine D2 and GABAB in the ovary of treated ticks.
Subject(s)
Acaricides , Pyrethrins , Rhipicephalus , Acaricides/pharmacology , Animals , Chromatin , Dopamine/pharmacology , Female , Pyrethrins/pharmacology , gamma-Aminobutyric Acid/pharmacologyABSTRACT
To combat the problem of acaricide resistance in ticks, efforts have been made to develop eco-friendly herbal acaricides which are safe for animal use with no possibilities of evoking environment and residual toxicity. In the present study, the acaricidal properties of a commonly available plant, Annona squamosa have been established to develop a novel antitick phytoformulation using leaf extract, suitable solvents and emulsifier for managing resistant tick infestations in animals. The quality control of the safe and stable phytoformulation using Rutin as a major constituent was maintained by HPTLC profiling. Several phytformulations (A-E) were prepared and formulation 'C' is identifried as the most effective showing 52.5-75% antitick activity against in vitro treated resistant strains of R. microplus (IVRI-IV and V) with 33.8-40.2% inhibition of oviposition while 65-85% mortality against multiacaricide resistant field populations. The phytoformulation provided 70.4% efficacy after first larval challenge in experimentally infested animals. The field efficacy of the formulation in different multilocational field trials was 68.53-77.8%. The detail antitick efficacy of the formulation along with safety and stability are explained as a component of antitck technology to manage resistant tick problem in livestock.
Subject(s)
Acaricides , Annona , Cattle Diseases , Plant Extracts , Rhipicephalus , Tick Infestations , Acaricides/pharmacology , Animals , Annona/chemistry , Cattle , Drug Resistance , Female , Larva , Plant Extracts/pharmacology , Tick Infestations/drug therapy , Tick Infestations/veterinaryABSTRACT
Prostaglandins are a group of important cell-signaling molecules involved in the regulation of ovarian maturation, oocyte development, egg laying and associated behaviors in invertebrates. However, the presence of prostaglandin E2 (PGE2), the key enzymes for PGE2 biosynthesis and its interference by drugs were not investigated previously in the ovary of ticks. The present study was undertaken to assess the modulation of the PGE2-mediated pathway in the eclosion blocking effect of flumethrin and terpenoid subfraction isolated from Artemisia nilagirica in Rhipicephalus annulatus ticks. The acaricidal activities and chemical profiling of the terpenoid subfraction were performed. The localization of the cyclooxygenase1 (COX1) and prostaglandin E synthase (PGES) enzymes and the quantification of PGE2 in the ovaries of the ticks treated with methanol (control), flumethrin and terpenoid subfraction were also undertaken. In addition, the vitellogenin concentration in hemolymph was also assayed. Both flumethrin and the terpenoid subfraction of A. nilagirica elicited a concentration-dependent inhibition of fecundity and blocking of hatching of the eggs. The COX1 could not be detected in the ovaries of treated and control ticks, while there was no significant difference observed in the concentration of vitellogenin (Vg) in them. The presence of PGES in the oocytes of control ticks was confirmed while the immunoreactivities against PGES were absent in the vitellogenic oocytes of ticks treated with flumethrin and terpenoid subfraction. The levels of PGE2 were below the detection limit in the ovaries of the flumethrin-treated ticks, while it was significantly lower in the ovaries of the terpenoid subfraction-treated ticks. Hence, the prostaglandin E synthase and PGE2 were identified as very important mediators for the signaling pathway for ovarian maturation and oviposition in ticks. In addition, the key enzyme for prostaglandin biosynthesis, PGES and the receptors for PGE2 can be exploited as potential drug targets for tick control. The detection of PGES by immunohistochemistry and quantification of PGE2 by LC-MSMS can be employed as valuable tools for screening newer compounds for their eclosion blocking acaricidal effects.
Subject(s)
Artemisia/chemistry , Dinoprostone/metabolism , Pyrethrins/pharmacology , Rhipicephalus/drug effects , Terpenes/isolation & purification , Terpenes/pharmacology , Animals , Antibodies/metabolism , Female , Gas Chromatography-Mass Spectrometry , Hemolymph/metabolism , Immersion , Ovary/drug effects , Ovary/enzymology , Peroxidase/metabolism , Prostaglandin-E Synthases/metabolism , Vitellogenins/metabolismABSTRACT
Animal production has a key role in global economic development and food security. Ticks, specifically Rhipicephalus microplus cause substantial economic and health impacts on more than eighty percent of the world cattle population. Though synthetic acaricides play a major role in tick management, their injudicious usage has caused environmental pollution and also promote the establishment of multi-acaricide resistant tick populations which is a matter of great concern. To provide an effective tool for controlling these resistant ticks, the present work was aimed to develop safe and inexpensive antitick natural formulations. Our bioprospection studies of Ageratum conyzoides plant established it as a species potentially having strong acaricidal activity due to the presence of potent acaricidal phyto-chemicals. To develop a suitable antitick natural formulation, 41 samples/fractions/formulations were prepared from the dry powder of the whole aerial part of the A. conyzoides plant using different techniques and delivery matrices. The strongest antitick effect was recorded for formulation ACF6, which demonstrated 87 ± 6% mean mortality with 57 % inhibition of oviposition in treated female ticks. Ticks treated with the ACF6 formulation showed a significant (p < 0.001) reduction in cuticular protein (1.238 ± 0.01 mg/mL) as compared to control ticks (2.928 ± 0.01 mg/mL) but no significant difference in chitin content of treated ticks and control ticks was observed. The formulation was found safe in a rat model as no significant differences in biochemical and haematological parameters among treated and control rats were noted. Histopathological studies indicated no sign of hepatocellular necrosis and no significant changes in the weights of liver and spleen was recorded. The overall in vivo efficacy of the formulation was 85 % for experimentally infested cattle with direct mortality of more than 80 % within 96 h post-application. The lethal effect of the formulation was in the form of drying and dead ticks 1-2 d after application. The developed formulation has the potential to be adopted as an alternative tick control measure in an ecofriendly manner.
Subject(s)
Acaricides , Ageratum/chemistry , Cattle Diseases/prevention & control , Drug Resistance , Plant Extracts , Rhipicephalus , Tick Control , Tick Infestations/veterinary , Animals , Cattle , Cattle Diseases/parasitology , Female , Larva/drug effects , Larva/growth & development , Male , Rhipicephalus/drug effects , Rhipicephalus/growth & development , Tick Infestations/parasitology , Tick Infestations/prevention & controlABSTRACT
The Hyalomma species of ticks have gained additional attention due to their role in the transmission of Theileria annulata infection in animals and the Crimean-Congo Haemorrhagic Fever (CCHF) virus in humans. Apart from these, many other pathogens viz., other species of Theileria, a few species of Babesia, Rickettsia and viruses are either maintained or transmitted by this tick species. The medium to large size species with longer proboscis has inflicted additional burden on the overall impact of tick infestations. Being a multi-host species, management of the species is very challenging. Presently, the traditional method of tick management using chemical acaricides is found insufficient and unsustainable. Henceforth, the overall burden of tick infestations and tick-borne diseases are increasing gradually. After the successful development of vaccines against cattle tick, Rhipicephalus microplus, the anti-Hyalomma vaccine is considered a feasible and sustainable management option. In the recent past research on herbal acaricides and its possible application for tick control seems promising. Other eco-friendly methods are still under experimental stage. The present review is focused on impact of Hyalomma species infestation on human and animal health with special emphasis on progress on its sustainable management.
ABSTRACT
Pu.1 is an ETS family transcription factor (TF) that plays critical roles in erythroid progenitors by promoting proliferation and blocking terminal differentiation. However, the mechanisms controlling expression and down-regulation of Pu.1 during early erythropoiesis have not been defined. In this study, we identify the actions of Runx1 and Pu.1 itself at the Pu.1 gene Upstream Regulatory Element (URE) as major regulators of Pu.1 expression in Burst-Forming Unit erythrocytes (BFUe). During early erythropoiesis, Runx1 and Pu.1 levels decline, and chromatin accessibility at the URE is lost. Ectopic expression of Runx1 or Pu.1, both of which bind the URE, prevents Pu.1 down-regulation and blocks terminal erythroid differentiation, resulting in extensive ex vivo proliferation and immortalization of erythroid progenitors. Ectopic expression of Runx1 in BFUe lacking a URE fails to block terminal erythroid differentiation. Thus, Runx1, acting at the URE, and Pu.1 itself directly regulate Pu.1 levels in erythroid cells, and loss of both factors is critical for Pu.1 down-regulation during terminal differentiation. The molecular mechanism of URE inactivation in erythroid cells through loss of TF binding represents a distinct pattern of Pu.1 regulation from those described in other hematopoietic cell types such as T cells which down-regulate Pu.1 through active repression. The importance of down-regulation of Runx1 and Pu.1 in erythropoiesis is further supported by genome-wide analyses showing that their DNA-binding motifs are highly overrepresented in regions that lose chromatin accessibility during early erythroid development.
Subject(s)
Cell Differentiation/genetics , Core Binding Factor Alpha 2 Subunit/genetics , Core Binding Factor Alpha 2 Subunit/metabolism , Erythroid Precursor Cells/cytology , Erythroid Precursor Cells/metabolism , Gene Expression Regulation , Proto-Oncogene Proteins/genetics , Trans-Activators/genetics , Animals , Chromatin/genetics , Chromatin/metabolism , Erythropoiesis/genetics , Mice , RAW 264.7 Cells , Response Elements , Transcription, GeneticABSTRACT
In the present study, the cytotoxic effects of amitraz, an octopamine receptor agonist on the reproductive system of engorged adult females of Rhipicephalus (Boophilus) annulatus were assessed using histology, electron microscopy and octopamine beta (OCTß) receptor transcriptional expression analysis. Adult immersion test (AIT) was performed by immersing the fully engorged female ticks for 2â¯min in different concentrations of amitraz (200, 250, 300, 350â¯ppm). Amitraz at the dose of 300â¯ppm, caused an adult tick mortality of 16.66⯱â¯6.80 per cent, inhibition of fecundity of 75.80 per cent and hatching of 50 per cent of ova laid by treated ticks. Histological changes in the ovaries of ticks collected after 24â¯h of treatment with amitraz (300â¯ppm), in comparison with controls (distilled water/methanol) were identified by microscopical examination of sections (4â¯â¯µm) stained using haematoxylin and eosin. These changes included reduction in size and basophilia of stage I oocytes, presence of cytoplasmic vacuoles of various sizes around germinal vesicle of stage II oocytes, wavy basement membrane of stage III oocytes and reduction in size and number of mature stage IV and V oocytes. Electron microscopy was employed for understanding the structural changes in the ultrathin sections (60â¯nm) of ovaries. Ticks treated with amitraz showed major ultrastructural changes such as irregular nuclear membrane, crystolysis of mitochondria and detachment of external and internal layers of basal lamina of oocytes. The cDNA synthesized from the total RNA of whole ticks and ovaries of ticks treated with amitraz along with controls were used for relative quantification of Octopamine ß receptor (OCTß-R) expression based on the 2-ΔΔCT method by quantitative real time PCR (qRT PCR). Glyceraldehyde-3-phosphate dehydrogenase (GAPDH) was used as endogenous control. Down regulation of expression of OCTß-R mRNA in the ovaries of amitraz treated ticks was observed compared to controls. Thus, the inhibition of fecundity observed in the ticks treated with amitraz can be attributed to the major structural changes and decreased expression of OCT ß receptor mRNA induced by it in the ovary.
Subject(s)
Insecticides/pharmacology , Rhipicephalus/drug effects , Toluidines/pharmacology , Analysis of Variance , Animals , Basement Membrane/drug effects , Basement Membrane/ultrastructure , Down-Regulation , Female , Fertility/drug effects , Gene Expression , Microscopy, Electron, Transmission , Mitochondria/drug effects , Mitochondria/ultrastructure , Oocytes/drug effects , Oocytes/ultrastructure , Ovary/anatomy & histology , Ovary/drug effects , Ovary/ultrastructure , Oviposition/drug effects , RNA, Messenger/isolation & purification , RNA, Messenger/metabolism , Real-Time Polymerase Chain Reaction , Receptors, Biogenic Amine/agonists , Receptors, Biogenic Amine/drug effects , Rhipicephalus/anatomy & histology , Rhipicephalus/genetics , Rhipicephalus/ultrastructure , Spectrophotometry , Tick Control/methods , Vacuoles/drug effects , Vacuoles/ultrastructureABSTRACT
BACKGROUND: We aimed to focus on the ixodid ticks parasitizing wild mammals and reptiles from Wayanad Wildlife Sanctuary, Western Ghat, southern India. METHODS: The taxonomic identification of ticks collected from wild mammals and reptiles was performed based on the morphology of adults. RESULTS: We revealed eight species of ticks including, Amblyomma integrum, Rhipicephalus (Boophilus) annulatus, Haemaphysalis (Kaiseriana) spinigera, H. (K.) shimoga, H. (K.) bispinosa, H. (Rhipistoma) indica, Rhipicephalus haemaphysaloides and R. sanguineus s.l. collected from nine species of wild mammals while four tick species Ablyomma kraneveldi, A. pattoni, A. gervaisi and A. javanense parasitizing on four species of reptiles. The highest host richness was shown by H. (K.) bispinosa and R. haemaphysaloides parasitizing six and five different host species, respectively. Reports of R. (B.) annulatus on sambar deer, A. javanense and A. kraneveldi on python as well as A. pattoni on Indian rat snake are the new host records from this region. CONCLUSION: Eight species of ticks parasitizing on nine species of wild mammals and four species of parasitizing on four species of reptiles were identified. The highest host richness was shown by H. (K.) bispinosa and R. haemaphysaloides. H. spinigera as the vector of KFD was also identified in this study.
ABSTRACT
The present study compares the in vitro efficacy of four chemical acaricides, viz. amitraz, coumaphos, deltamethrin and lindane, against Rhipicephalus (Boophilus) annulatus and Haemaphysalis bispinosa ticks based on adult immersion tests. Amitraz, at 350 ppm, elicited 29.2 ± 4.17% mortality against R. (B.) annulatus, 100% inhibition of fecundity and absence of hatching of eggs laid by treated ticks. The same compound at 300 ppm caused 62.5 ± 12.5% mortality against H. bispinosa, 96.7% inhibition of fecundity and complete blocking of eclosion. The LC50 value of amitraz against susceptible H. bispinosa was 181 ppm. Deltamethrin at 400 ppm, elicited 25.0 ± 4.81% adult R. (B.) annulatus mortality, 97.5% inhibition of fecundity and absence of egg hatching. Complete blocking of egg hatching was observed even at 30 ppm. However, deltamethrin (at 50 ppm) elicited 75.0 ± 10.76% mortality against H. bispinosa, 65.8% inhibition of fecundity and very low egg hatching (10%). The LC50 for deltamethrin against susceptible H. bispinosa was 33.8 ppm. Coumaphos at 50 ppm, caused mortality of 70.8 ± 4.17% with R. (B.) annulatus whereas 100% mortality was observed against H. bispinosa. The LC50 values of coumaphos against R. (B.) annulatus and H. bispinosa were 9 and 8.75 ppm, respectively. Complete inhibition (100%) of fecundity was observed even at 30 ppm against both parasites. Complete blocking of egg hatching was also observed even at 10 ppm of coumaphos. Lindane at 1000 ppm caused mortality of 87.5 ± 7.98% against R. (B.) annulatus and 83.3% mortality against H. bispinosa at 100 ppm. The LC50 values of lindane against R. (B.) annulatus and H. bispinosa were 157 and 8.61 ppm, respectively. Complete inhibition of fecundity was observed with R. (B.) annulatus treated with lindane above 200 ppm and with H. bispinosa at a concentration above 50 ppm. Complete blocking of egg hatching was observed in R. (B.) annulatus, even at 100 ppm. Lindane caused 100% blocking of egg hatching at 1 ppm in the case of H. bispinosa.
Subject(s)
Acaricides , Ixodidae , Tick Control , Animals , Coumaphos , Female , Hexachlorocyclohexane , In Vitro Techniques , Nitriles , Pyrethrins , Rhipicephalus , Species Specificity , ToluidinesABSTRACT
Sustained and controlled release of neurotrophic factors in target tissue through nanomaterial based delivery system could be a better strategy for nerve tissue regeneration. The present study aims to prepare the nerve growth factor (NGF) encapsulated chitosan nanoparticles (NGF-CNPs) and its evaluation on neuronal differentiation potentiality of canine bone marrow derived mesenchymal stem cells (cBM-MSCs). The NGF-CNPs were prepared by ionotropic gelation method with tripolyphosphate (TPP) as an ionic cross-linking agent. Observations on physiochemical properties displayed the size of nanoparticles as 80-90 nm with positive zeta potential as well as an ionic interaction between NGF and nanoparticle. NGF loading efficiency was found to be 61% while its sustained release was observed by an in vitro release kinetics study. These nanoparticles were found to be cytocompatible to cBM-MSCs when supplemented at a concentration upto 4 mg/ml in culture media. The NGF-CNP supplemented culture media was able to transdifferentiate the preinduced cBM-MSCs into neurons in a better way than unbound NGF supplementation. Further, it was also noticed that NGF-CNPs were able to transdifferentiate cBM-MSCs without any chemical based preinduction. In conclusion, our findings propose that NGF-CNPs are capable of releasing bioactive NGF with the ability to transdifferentiate mesenchymal stem cells into neurons, suggesting its potential future application in nerve tissue regeneration.
Subject(s)
Chitosan/chemistry , Mesenchymal Stem Cells/cytology , Metal Nanoparticles/chemistry , Nerve Growth Factor/chemistry , Neurons/cytology , Animals , Apoptosis , Biological Assay , Cell Differentiation , Cell Proliferation , Cross-Linking Reagents/chemistry , Culture Media , Dogs , Drug Delivery Systems , Equipment Design , Flow Cytometry , Ions , Nanoparticles/chemistry , Nerve Regeneration , Neurons/drug effects , Polyphosphates/chemistryABSTRACT
The present study utilizes the ultrastructural analysis of the fully engorged female Rhipicephalus (Boophilus) annulatus ticks, as a tool to evaluate the cytotoxic potential of deltamethrin and amitraz on the germinative cells. The ultrastructural analysis of the ovary of the normal (untreated) R (B.) annulatus revealed, oocytes in different stages of development, attached to the ovary wall by pedicel cells. The attachment site of oocyte to the pedicel cell was characterized by indentations of the plasma membrane. The oocyte was bound by three cell membranes viz., plasma membrane, chorion and basal lamina. The stages of oocytes were differentiated ultrastructurally based on the features of their outer membrane and the number and size of lipid and yolk droplets. Detailed day wise analysis of ultrastructural changes in the ovary during the post-engorgement period revealed the occurrence of the degenerative changes from day five onwards. These appeared first in the oocytes followed by the germinal epithelium. The ovary of ticks treated with methanol (control), revealed similar topographies as that of a normal ovary except for the presence of very few oocytes with ring shaped nucleoli. Ultrastructurally, treatment with deltamethrin produced more prominent and extensive morphological alterations when compared to amitraz. In the case of ticks treated with amitraz, the oocytes of stage IV and V showed wavy and disrupted outer boundaries along with the loss of integrity of the yolk droplets. Uneven nuclear membranes of stage II oocytes and cristolysis of mitochondria of mature oocytes were the other changes noticed. Ticks treated with deltamethrin revealed prominent modifications such as, detachment of the basal lamina, wrinkled boundary, inconsistent nuclear membrane, ring shaped nucleoli and chromatin clumping in the case of the early stage oocytes (I and II), whereas swelling and cristolysis of mitochondria were seen in mature oocytes. The study further indicated that, in addition to the previous proven neurotoxic effects, these compounds act directly on the ovary of tick.
Subject(s)
Acaricides/pharmacology , Nitriles/pharmacology , Pyrethrins/pharmacology , Rhipicephalus/ultrastructure , Tick Infestations/veterinary , Toluidines/pharmacology , Animals , Female , Microscopy, Electron, Transmission/veterinary , Oocytes/drug effects , Oocytes/ultrastructure , Ovary/drug effects , Ovary/ultrastructure , Rhipicephalus/drug effects , Tick Infestations/drug therapy , Tick Infestations/parasitologyABSTRACT
In the present study, the acaricidal effects of caffeic acid, nortriptyline, precocene I and quercetin against Rhipicephalus annulatus (syn. Boophilus annulatus) Say (Acari: Ixodidae) were evaluated. Adult immersion technique (24 ticks immersed for 2 min in one dilution of the compound) was used for the assessment of the effects of caffeic acid (0.39-100 mg/mL), nortriptyline (0.625-50 mg/L), precocene I (0.004488-5 mg/mL) and quercetin (6.25-100 mg/mL) against R. annulatus. Adult tick mortality, reproductive index, inhibition of fecundity and hatching were calculated. Caffeic acid, nortriptyline, precocene I and quercetin revealed very low adult mortality and inhibition of fecundity, even at the highest concentration tested. Quercetin (>50 mg/mL) caused blocking of hatching of eggs.
Subject(s)
Acaricides/pharmacology , Plant Extracts/pharmacology , Rhipicephalus/drug effects , Animals , Female , Ovum/drug effectsABSTRACT
To identify suitable targets for development of cross-protective tick vaccine, in silico analysis was attempted and male tick derived molecule, voraxin-α was targeted. The voraxin-α homologue of Rhipicephalus (Boophilus) microplus was cloned, sequenced and analyzed employing standard methods. The deduced amino acids sequence analysis of the 419 bp cloned voraxin-α gene of R. (B.) microplus indicated very high (94.6 %) similarity with voraxin-α of the R. appendiculatus and moderate to low identity with Amblyomma hebraeum, Dermacentor silvarum and Haemaphysalis longicornis. The results suggest that recombinant voraxin-α might be a good candidate as cross-protective anti-tick vaccine.
ABSTRACT
To develop an eco-friendly tick control method, seven plant extracts were prepared using 50 and 95% ethanol and evaluated for acaricidal activity against cattle tick, Rhipicephalus (Boophilus) microplus. The adult immersion test was adopted for testing different extracts. Based on 72 h screening criterion, 95% ethanolic extracts of Datura metel fruits and Argemone mexicana whole plant were found effective showing more than 50% mortality of treated ticks. The 95% ethanolic extracts of D. metel fruits and A. mexicana whole plant exhibited acaricidal and reproductive inhibitory effects on treated ticks. The LC90 values of D. metel and A. mexicana extracts were determined as 7.13 and 11.3%, respectively. However, although both the extracts were found efficacious against deltamethrin-resistant IVRI-4 and multi-acaricide resistant IVRI-5 lines of R. (B.) microplus, they caused less mortality than treated ticks of the reference IVRI-I line. Phytochemical studies indicated the presence of alkaloids and glucosides in D. metel fruits and alkaloids, terpenoids, flavonoids and phenolics in A. mexicana whole plant extracts. The results indicated that these botanicals may play an important role in reducing the use of chemicals for tick control and possibly to manage resistant tick population in environment friendly manner.
Subject(s)
Magnoliopsida/chemistry , Plant Extracts/pharmacology , Rhipicephalus/drug effects , Tick Control/methods , Acaricides/pharmacology , Animals , Cattle , Cattle Diseases/parasitology , Cattle Diseases/prevention & control , Drug Resistance , Female , Tick Infestations/parasitology , Tick Infestations/prevention & control , Tick Infestations/veterinaryABSTRACT
The present communication describes the detailed day wise study of histological changes of the ovary of Rhipicephalus (Boophilus) annulatus in the postengorgement period together with the systematic classification of their oocytes. The ovary of R. (B.) annulatus is panoistic type with an asynchronous development of oocytes. All the stages (II, III, IV, and V) of oocytes except stage I were similar to R. (B.) microplus. The stage I oocytes showed basophilia, which was not reported earlier in other species of ticks. Day wise changes were in the form of presence of oogonia in partially fed and day one engorged adults, considerable degeneration of oocytes on day two, emergence of new wave of oocytes on day three, presence of mature oocytes up to day eight, and complete degeneration of ovarian tissue from day eight onwards. The degenerative changes in the ovary appeared initially in the oocytes followed by germinal epithelium.